New Technologies for Patch-Clamp Recording

描述：膜片钳技术是现代医学的核心技术。 细胞电生理学。它用于获取有关以下内容的明确信息 离子通道和转运体中的门控和离子传输机制，以及 提供了一种将配体和调节剂应用于这些膜的方法 蛋白质，同时观察它们的功能。膜片钳技术也是 首屈一指的单分子技术，其中一个随机行为 可以直接观察到单个离子通道。 我们建议应用一项有关电极材料的令人兴奋的新发现 和制造技术，极大地提高了灵敏度和 制作膜片钳记录的简便性。在拟议的工作中，我们 将应用微制造技术来提炼一种新的 基于聚二甲基硅氧烷的微电极技术。PDMS是一种有机硅 可对其表面进行改性以与电池形成高阻密封的聚合物 膜；它也可以被微模塑成非常小的特征尺寸以形成 用于膜片钳大阵列的电极孔和细胞界面 电极。我们建议开发一个非常低噪声的膜片钳系统，该系统基于 PDMS电极技术，以及电极阵列，使 非常简单的“盲打”培养细胞。我们还建议将 微流控装置首次进入电极，允许快速 交换电极溶液。 这种记录技术的灵敏度的大幅提高将打开 作为单分子事件的新研究领域将在短得多的时间内被看到 时间尺度。拟议的膜片钳技术的简化将使 在寻找离子通道和转运蛋白基因以及 在发现新的药理制剂方面。

DESCRIPTION: The patch-clamp technique is the central technique of modern cellular electrophysiology. It is used to obtain definitive information about gating and ion-transport mechanisms in ion channels and transporters, and provides a way in which ligands and modulators can be applied to these membrane proteins while their function is observed. The patch-clamp technique is also the premier single-molecule technique, where the stochastic behavior of a single ion channel can be observed directly. We propose to apply an exciting new discovery concerning electrode materials and fabrication techniques to greatly increase both the sensitivity and the ease with which patch-clamp recordings can be made. In the proposed work, we will apply microfabrication technologies to the refinement of a novel microelectrode technology based on poly (dimethylsiloxane). PDMS is a silicone polymer whose surface can be modified to form high-resistance seals with cell membranes; it can also be micromolded to very small feature sizes to form electrode apertures and the cellular interface for large arrays of patch-clamp electrodes. We propose to develop a very low-noise patch-clamp system based on the PDMS electrode technology, as well as electrode arrays to make possible the very simple "blind" patching of cultured cells. We also propose to incorporate microfluidic devices into the electrodes to allow for the first time the rapid exchange of the electrode solution. A substantial increase in sensitivity of this recording technique would open new areas of research as single-molecule events will be visible on much shorter time scales. The proposed simplification of the patch-clamp technique will have far-reaching benefits in the search for ion channel and transporter genes and in the discovery of new pharmacological agents.